Subsurface imaging of silicon nanowire circuits and iron oxide nanoparticles with sub-10 nm spatial resolution

Non-destructive subsurface characterization of nanoscale structures and devices is of significant interest in nanolithography and nanomanufacturing. In those areas, the accurate location of the buried structures and their nanomechanical properties are relevant for optimization of the nanofabrication process and the functionality of the system. Here we demonstrate the capabilities of bimodal and trimodal force microscopy for imaging silicon nanowire devices buried under an ultrathin polymer film. We resolve the morphology and periodicities of silicon nanowire pairs. We report a spatial resolution in the sub-10 nm range for nanostructures buried under a 70 nm thick polymer film. By using numerical simulations we explain the role of the excited modes in the subsurface imaging process. Independent of the bimodal or trimodal atomic force microscopy approach, the fundamental mode is the most suitable for tracking the topography while the higher modes modulate the interaction of the tip with the buried nanostructures and provide subsurface contrast.This work was funded by the European Union FP7/2007-2013 under Grant Agreement No. 318804 (SNM), the Ministerio de Economía y Competitividad (Spain) under grants MAT2013-44858-R, CSD2010-00024 and the European Research Council ERC-AdG-340177 (3DNanoMech).Peer reviewe

TLR-Mediated Host Immune Response to Parasitic Infectious Diseases

Toll-like receptors (TLRs) are important for the host immune response to a variety of pathogens, including bacteria, viruses, fungi, and parasites. These receptors become activated upon recognizing pathogen-associated molecular patterns (PAMPs) and thus initiate the innate immune response to the corresponding pathogen. A key aspect of TLRs is their activation of signaling that leads to cytokine production and an inflammatory response. Additionally, TLRs act as the bridge between innate and acquired immunity, enhancing phagocytosis and the process of killing parasites. We herein focus on how parasites (protozoans and helminths) and their derived products have the capability of stimulating or evading the host response by triggering or inhibiting TLR activation. Parasites often develop successful survival strategies that imply interference with the host immune response. Accordingly, many of these organisms have molecules that modulate inflammation and other aspects of host immunity. Taking advantage of such mechanisms, there are some anti-inflammatory therapies based on human infection with helminths. Helminths and protozoans influence the activity of various TLRs, especially TLR2, TLR4, and TLR9. A better understanding of the role of TLRs and their parasite-derived ligands should certainly provide new therapeutic tools for combatting various parasitic and inflammatory diseases

Agenda de investigación y monitoreo en bosques secos de Colombia (2013-2015): fortaleciendo redes de colaboración para su gestión integral en el territorio

Tropical dry forests are the most threatened, yet among the least studied ecosystems in the Neotropics. These forests hold a unique combination of plants, animals and microorganisms that have adapted to the rain seasonality that characterizes tropical dry forest, thus holding many endemic species. In addition, this ecosystem provides key environmental services such as water regulation, soil retention and carbon sequestration, which regulates local climate, water and nutrients availability. In Colombia, dry forests occur in six regions: the Caribbean, the inter-Andean valleys of Cauca and Magdalena, the Northeastern Andes, the Patía valley, and the Llanos (Orinoquia region). In this working paper we discuss the three lines of research that the Instituto Alexander von Humboldt has been leading (2013-2015) to enhance tropical dry forest research, longtermmonitoring, and sustainable management: i) distribution and conservation status, ii) composition, ecological dynamics and functioning, and iii) characterization and conservation of tropical dry forests associated biodiversity.El bosque seco tropical tiene una diversidad única de plantas, animales y microorganismos que se han adaptado a condiciones de estrés hídrico, por lo cual presenta altos niveles de endemismo. Presta además servicios fundamentales como la regulación hídrica, la retención de suelos y la captura de carbono que regula el clima y la disponibilidad de agua y nutrientes. En Colombia el bosque seco se encuentra en seis regiones biogeográficas: el Caribe, los valles interandinos del Cauca y el Magdalena, la región Norandina en Santander y Norte de Santander, el valle del río Patía y afloramientos rocosos en los departamentos de Arauca y Vichada en los Llanos Orientales. Pese a su importancia para la conservación de la biodiversidad y los beneficios derivados para las sociedades humanas, este ecosistema es considerado el más amenazado del neotrópico y cuenta con muy poca información disponible, producto de la investigación científica. Bajo este contexto, el Instituto Humboldt, bajo el liderazgo de los Programas de Ciencias de la Biodiversidad y de Gestión Territorial, ha liderado la consolidación de una Agenda de Investigación y Monitoreo a escala de país para fortalecer la gestión integral de este ecosistema a partir del conocimiento científico. En este documento de trabajo se analizan las tres líneas de investigación que se han venido liderando desde 2013 hasta 2015 con el objetivo de fomentar la investigación, el monitoreo a largo plazo y un manejo integral del bosque seco en Colombia: i) distribución espacial y estado de conservación, ii) composición, dinámica y funcionamiento, y iii) caracterización y conservación de la biodiversidad asociada

Metabolic rates in Atlantic bluefin tuna larvae: first data and methodological challenges

Atlantic bluefin tuna is an emblematic scombrid species, but many physiological aspects during the larval stages are still unknown. The mechanisms of how fish larvae balance growth and activity are of great interest since metabolic costs are very high compared to juveniles and adults. However, there is a lack of information about metabolic costs in scombrid larvae. This lack of data is probably related to the challenges associated to larval handling before and during respirometry trials. In this study for the first time, we: i) estimate the relationship between routine metabolic rate and the larval dry weight (mass scaling exponent) at 26°C, ii) measure metabolism under light and darkness and iii) explore the influence of nutritional status (RNA:DNA ratio) on the inter-individual variability in metabolic rates. The relationship between metabolism and size (ranging from 0.6 to 23 mg) was near isometric (slope, b=0.99), in contrast to the allometric relationship observed in most species (b=0.87). Our results show no significant differences in oxygen consumption under light and darkness. A possible regulation (decrease) of their swimming activity with the consequent decrease in the oxygen consumption in light situations is discussed. Nutritional condition did not explain the inter-individual differences in oxygen consumption. This study first reports metabolic rates of Atlantic bluefin tuna larvae and discusses the challenges of performing bioenergetics studies with early life stages of Scombrids

Linear classifier and textural analysis of optical scattering images for tumor classification during breast cancer extraction

Texture analysis of light scattering in tissue is proposed to obtain diagnostic information from breast cancer specimens. Light scattering measurements are minimally invasive, and allow the estimation of tissue morphology to guide the surgeon in resection surgeries. The usability of scatter signatures acquired with a micro-sampling reflectance spectral imaging system was improved utilizing an empirical approximation to the Mie theory to estimate the scattering power on a per-pixel basis. Co-occurrence analysis is then applied to the scattering power images to extract the textural features. A statistical analysis of the features demonstrated the suitability of the autocorrelation for the classification of notmalignant (normal epithelia and stroma, benign epithelia and stroma, inflammation), malignant (DCIS, IDC, ILC) and adipose tissue, since it reveals morphological information of tissue. Non-malignant tissue shows higher autocorrelation values while adipose tissue presents a very low autocorrelation on its scatter texture, being malignant the middle ground. Consequently, a fast linear classifier based on the consideration of just one straightforward feature is enough for providing relevant diagnostic information. A leave-one-out validation of the linear classifier on 29 samples with 48 regions of interest showed classification accuracies of 98.74% on adipose tissue, 82.67% on non-malignant tissue and 72.37% on malignant tissue, in comparison with the biopsy H and E gold standard. This demonstrates that autocorrelation analysis of scatter signatures is a very computationally efficient and automated approach to provide pathological information in real-time to guide surgeon during tissue resection.This work has been supported by CYCIT projects DA2TOI (FIS2010-19860) and TFS (TEC2010-20224-C02-02), as well as FPU PhD Scholarship (FPU12/04130) funded by the Spanish Government

Saturated fatty acid-enriched small extracellular vesicles mediate a crosstalk inducing liver inflammation and hepatocyte insulin resistance

[Background & Aims]: Lipotoxicity triggers non-alcoholic fatty liver disease (NAFLD) progression owing to the accumulation of toxic lipids in hepatocytes including saturated fatty acids (SFAs), which activate pro-inflammatory pathways. We investigated the impact of hepatocyte- or circulating-derived small extracellular vesicles (sEV) secreted under NAFLD conditions on liver inflammation and hepatocyte insulin signalling. [Methods]: sEV released by primary mouse hepatocytes, characterised and analysed by lipidomics, were added to mouse macrophages/Kupffer cells (KC) to monitor internalisation and inflammatory responses. Insulin signalling was analysed in hepatocytes exposed to conditioned media from sEV-loaded macrophages/KC. Mice were i.v. injected sEV to study liver inflammation and insulin signalling. Circulating sEV from mice and humans with NAFLD were used to evaluate macrophage–hepatocyte crosstalk. [Results]: Numbers of sEV released by hepatocytes increased under NAFLD conditions. Lipotoxic sEV were internalised by macrophages through the endosomal pathway and induced pro-inflammatory responses that were ameliorated by pharmacological inhibition or deletion of Toll-like receptor-4 (TLR4). Hepatocyte insulin signalling was impaired upon treatment with conditioned media from macrophages/KC loaded with lipotoxic sEV. Both hepatocyte-released lipotoxic sEV and the recipient macrophages/KC were enriched in palmitic (C16:0) and stearic (C18:0) SFAs, well-known TLR4 activators. Upon injection, lipotoxic sEV rapidly reached KC, triggering a pro-inflammatory response in the liver monitored by Jun N-terminal kinase (JNK) phosphorylation, NF-κB nuclear translocation, pro-inflammatory cytokine expression, and infiltration of immune cells into the liver parenchyma. sEV-mediated liver inflammation was attenuated by pharmacological inhibition or deletion of TLR4 in myeloid cells. Macrophage inflammation and subsequent hepatocyte insulin resistance were also induced by circulating sEV from mice and humans with NAFLD. [Conclusions]: We identified hepatocyte-derived sEV as SFA transporters targeting macrophages/KC and activating a TLR4-mediated pro-inflammatory response enough to induce hepatocyte insulin resistance.This work was supported by grants PID2021-122766OB-I00 (AMV), PID2019-105989RB-I00 (JB), PID2020-113238RB-I00 (LB), PID2019-106581RB-I00 (MAM), PID2020-114148RB-I00 (MI), PID2019-107036RB-I00 (RF), and RD21/0006/0001 (ISCIII) (IL) funded by Ministerio de Ciencia e Innovación/Agencia Estatal de Investigación/10.13039/501100011033 and ERDF ‘A way of making Europe’ by the European Union (MICINN/AEI/FEDER, EU), grant EFSD/Boehringer Ingelheim European Research Programme on ‘Multi-System Challenges in Diabetes’ from the European Foundation for the Study of Diabetes (AMV), P2022/BMD-7227 (Comunidad de Madrid, Spain) (AMV), Fundación Ramón Areces (Spain) (AMV), CIBERdem (AMV and JB), CIBERhed (RF), and CIBERcv (LB) (ISCIII, Spain). LB and AMV belong to the Spanish National Research Council’s (CSIC’s) Cancer Hub. We also acknowledge the Spanish Ministry of Economy and Competitiveness (MINECO) postdoctoral contract IJCI-2015-24758 to IGM and the Spanish Ministry of Education, Culture and Sport (MECD) FPU17/02786 grant to RA

Comparative analysis of acoustic therapies for tinnitus treatment based on auditory event-related potentials

IntroductionSo far, Auditory Event-Related Potential (AERP) features have been used to characterize neural activity of patients with tinnitus. However, these EEG patterns could be used to evaluate tinnitus evolution as well. The aim of the present study is to propose a methodology based on AERPs to evaluate the effectiveness of four acoustic therapies for tinnitus treatment.MethodsThe acoustic therapies were: (1) Tinnitus Retraining Therapy (TRT), (2) Auditory Discrimination Therapy (ADT), (3) Therapy for Enriched Acoustic Environment (TEAE), and (4) Binaural Beats Therapy (BBT). In addition, relaxing music was included as a placebo for both: tinnitus sufferers and healthy individuals. To meet this aim, 103 participants were recruited, 53% were females and 47% were males. All the participants were treated for 8 weeks with one of these five sounds, which were moreover tuned in accordance with the acoustic features of their tinnitus (if applied) and hearing loss. They were electroencephalographically monitored before and after their acoustic therapy, and wherefrom AERPs were estimated. The sound effect of acoustic therapies was evaluated by examining the area under the curve of those AERPs. Two parameters were obtained: (1) amplitude and (2) topographical distribution.ResultsThe findings of the investigation showed that after an 8-week treatment, TRT and ADT, respectively achieved significant neurophysiological changes over somatosensory and occipital regions. On one hand, TRT increased the tinnitus perception. On the other hand, ADT redirected the tinnitus attention, what in turn diminished the tinnitus perception. Tinnitus handicapped inventory outcomes verified these neurophysiological findings, revealing that 31% of patients in each group reported that TRT increased tinnitus perception, but ADT diminished it.DiscussionTinnitus has been identified as a multifactorial condition highly associated with hearing loss, age, sex, marital status, education, and even, employment. However, no conclusive evidence has been found yet. In this study, a significant (but low) correlation was found between tinnitus intensity and right ear hearing loss, left ear hearing loss, heart rate, area under the curve of AERPs, and acoustic therapy. This study raises the possibility to assign acoustic therapies by neurophysiological response of patient